RAPID LOW COST DNA SEQUENCING BY MASS SPECTROMETRY

通过质谱法进行快速、低成本的 DNA 测序

• 批准号: 3333674
• 负责人: ROBERT J LEVIS
• 金额: $ 21.77万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3333674
• 关键词: DNA; biomedical automation; biomedical equipment development; genetic techniques; health economics; mass spectrometry; nucleic acid structure

The goal of this proposal is the development of a novel mass spectral method which will permit the sequencing of 500 base-long (or longer) strands of DNA. The realization of this instrument will allow DNA sequencing to be performed at a rate approaching one million bases per day- a rate three orders faster than currently available techniques. To achieve this objective, we plan on melding current DNA sequencing strategies with three new technologies to produce a low-cost, easily automated sequencing method. We intend (i) to draw on our expertise in laser vaporization to develop a new vaporization strategy that will permit precise control over the chemical nature of the desorbed DNA and will be very amenable to rapid sequencing; (ii) to utilize an ionization scheme that will allow selective ionization of the vaporized DNA strands and ultrasensitive detection of these strands; and (iii) to analyze these ionized DNA strands by a mass spectral technique that will have the sensitivity and resolution required to sequence DNA up to 1000 bases long. The automation of the technology we propose would result in an extremely sensitive DNA sequencing method requiring little manual labor. The sequencing cost resulting from an instrument that used this methodology should be less than $0.10 per sequenced base, thus resulting in at least a 10-fold reduction in the sequencing costs associated with the Human Genome Initiative.

该提案的目标是开发一种新的质谱 该方法将允许测序500个碱基长（或更长）的 DNA链 这个仪器的实现将使DNA 测序速度接近每天一百万个碱基- 比目前可用的技术快三个数量级。 实现 为此目标，我们计划将当前的DNA测序策略与 三项新技术，以产生低成本，易于自动化测序 法 我们打算（i）利用我们在激光汽化方面的专业知识， 开发一种新的汽化策略， 解吸附的DNA的化学性质，并且将非常适合于快速分析。 测序;（ii）利用电离方案， 蒸发的DNA链的电离和超灵敏的检测 这些链;和（iii）通过质量分析这些电离的DNA链， 具有所需灵敏度和分辨率的光谱技术 能对1000个碱基的DNA进行测序 自动化的技术，我们 这一提议将导致一种极其灵敏的DNA测序方法 需要很少的体力劳动。 由一个 使用这种方法的工具应低于0.10美元， 测序的碱基，从而导致至少10倍的减少， 与人类基因组计划相关的测序成本。